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Eugene Higgins Professor of Cell Biology
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Phone: (203) 737-4461 Lab: (203) 737-4457/-4469 Fax: (203) 737-4436 e-mail: pietro.decamilli@yale.edu |
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Department of Cell Biology <Courier Address> |
A main focus of our research is the elucidation of the mechanisms responsible for the biogenesis and traffic of synaptic vesicles, the secretory organelles that store and secrete fast-acting neurotransmitters. Synaptic vesicles deliver their content into the synaptic space by fusion with the plasma membrane (exocytosis) and are rapidly reformed by the endocytosis and recycling of their membranes. Thus, studies of these organelles have general relevance for the understanding of mechanisms involved in the secretory and endocytic pathways in all cells. Current projects address 1) the clathrin-dependent internalization of synaptic vesicles, 2) the mechanisms underlying acquisition of membrane curvature and vesicle fission, 3) the biochemical steps involved in the transformation of an uncoated clathrin coated vesicle into a mature synaptic vesicle, 4) the role of the cortical cytoskeleton in regulating synaptic vesicle traffic to and from the plasma membrane. We are particularly interested in the role of protein-lipid interactions in vesicle traffic. These studies have led us to discover an important function of inositol phospholipids (phosphoinositides) in synaptic vesicle recycling and we are actively pursuing studies on the regulatory function of these phospholipids at synapses.
Click for QuickTime movie (4.28 MB)
Figure 1. Detection of phosphatidylinositolphosphate by ESI/MS-MS. Product scan of the material generated by fragmentation of the ion at m/z 885.7. The peaks obtained correspond to the expected fragments of phosphatidylinositol (from the work of Markus Wenk and Pietro De Camilli).
Figure 2. Immunogold labeling for PIP kinase type I
of cell free reactions in which synaptic membranes were incubated
with brain cytosol, ATP and GTPS to induce the formation of clathrin coated intermediates. Note the concentration of PIP
kinase type I, a PI(4)P 5-kinase, on the membrane from which coated pits (arrows) originate, in agreement with the known
interaction of clathrin adaptors with PI(4,5)P2 (from the work of Markus Wenk, Lorenzo Pellegrini, Gilbert Di Paolo and
Pietro De Camilli)
Figure 3. Electron micrograph of a synapse in the cerebellar cortex.
Figure 4. A cycle of PI(4,5)P2 synthesis and hydrolysis accompanies the exo-endocytosis of synaptic vesicles at synapses.
Selected Publications:
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for PDF
Lee SY, Voronov S, Letinic K, Nairn AC, Di Paolo G, and De Camilli P. (2005) Regulation of the interaction between phosphatidylinositol(4)phosphate 5-kinase type I
and talin by proline-directed protein kinases. J Cell Biol. 8(5):789-99.
Chen H, De Camilli P. (2005) The association of epsin with ubiquitinated cargo along the endocytic pathway is negatively regulated by its interaction with clathrin. Proc Natl Acad Sci USA. 102(8): 2766-2771.
Cestra G, Toomre D, Chang S, De Camilli P. (2005) The Abl/Arg substrate ArgBP2/nArgBP2 coordinates the function of multiple regulatory mechanisms converging on the actin cytoskeleton. Proc Natl Acad Sci USA. 102(5): 1731-1736.
Morgan JR, Di Paolo G, Werner H, Shchedrina VA, Pypaert M, Pieribone VA, De Camilli P. (2004) A role for talin in presynaptic function. J Cell Biol. 167(1):43-50.
Di Paolo G, Moskowitz HS, Gipson K, Wenk MR, Voronov S, Obayashi M, Flavell R, Fitzsimonds RM, Ryan TA, De Camilli P. (2004) Impaired PtdIns(4,5)P2 synthesis in nerve terminals produces defects in synaptic vesicle trafficking. Nature 431(7007):415-22.
Wenk MR, De Camilli P. (2004) Protein-lipid interactions and phosphoinositide metabolism in membrane traffic: insights from vesicle recycling in nerve terminals. Proc Natl Acad Sci USA. 101(22): 8262-9.
Wenk MR, Lucast L, Di Paolo G, Romanelli AJ, Suchy SF, Nussbaum RL, Cline GW, McMurray W and De Camilli P. (2003) Phosphoinositide profiling in complex lipid mixtures using electrospray ionization mass spectrometry. Nat Biotech. 21: 813-817.
Murthy V. and De Camilli P. (2003) Cell Biology of the presynaptic terminal. Ann. Rev. Neurosci. 26: 701-728.
Di Paolo G, Pellegrini L, Letinic K, Voronov S, Chang S, Wenk MR, and De Camilli P. (2002) Recruitment and regulation of PIP kinase type I by the FERM domain of talin. Nature. 420: 85-89.
Lee E, Marcucci M, Daniell L, Ochoa GC, Farsad K, and De Camilli P. (2002) Amphiphysin 2 (Bin1) and T-tubule biogenesis in muscle. Science. 297: 1193-1196.
Polo S, Bossi G, Sigismund S, Feretta M, Guidi M, Capuo MR, Bossi G, Chen H, De Camilli P, and Di Fiore PP. (2002) A single motif responsible for ubiquitin recognition and monoubiquitation in endocytic proteins. Nature. 416: 451-455.
